In vitro pre-conditioned bone marrow-derived mesenchymal stem cells and uses thereof

Abstract

Disclosed is a composition including: an isolated in vitro pre-conditioned population of adult bone marrow derived mesenchymal stem cells (BMSCs), wherein the BMSCs express neuronal markers, and wherein the neuronal markers are PGP9.5, NSE, Tuj1, HuC/D and neuronal nitric oxide synthase (nNOS). Methods of preparing the BMSCs are also provided. In addition, the present disclosure is directed to a method of treating an enteric nervous system-related disorder including: administering to a subject in need thereof a pharmaceutical composition including the in vitro pre-conditioned BMSC population and a pharmaceutically acceptable carrier.

What is claimed is: 
     
       1. A composition comprising:
 an isolated in vitro pre-conditioned population of adult bone marrow derived mesenchymal stem cells (BMSCs),
 wherein at least 80% of the in vitro pre-conditioned BMSC population express PGP9.5 and NSE, 
 wherein at least 70% of the in vitro pre-conditioned BMSC population express Tuj1 and HuC/D, 
 wherein at least 75% of the in vitro pre-conditioned BMSC population express neuronal nitric oxide synthase (nNOS), and 
 wherein the BMSCs promote de novo regeneration of neurons not originated from the BMSCs when the BMSCs are administered to a subject. 
 
 
     
     
       2. The composition of  claim 1 , wherein the in vitro pre-conditioning comprises:
 providing a population of mesenchymal stem cells from a bone marrow; 
 culturing the population of mesenchymal stem cells in a medium comprising a glial cell derived neurotrophic factor and a fetal gut culture medium. 
 
     
     
       3. The composition of  claim 1 , wherein about 80% of the cells in the in vitro pre-conditioned BMSC population express PGP9.5, about 80% of the cells in the in vitro pre-conditioned BMSC population express NSE, about 75% of the cells in the in vitro pre-conditioned BMSC population express Tuj1, about 73% of the cells in the in vitro pre-conditioned BMSC population express HuC/D and about 75% of the in vitro pre-conditioned BMSC population express nNOS. 
     
     
       4. The composition of  claim 1 , wherein the in vitro pre-conditioned BMSCs are capable of maintaining a neuronal-like phenotype in vivo for at least 28 days. 
     
     
       5. The composition of  claim 1 , wherein the in vitro pre-conditioned BMSCs are human cells. 
     
     
       6. The composition of  claim 1 , further comprising a pharmaceutically acceptable carrier. 
     
     
       7. The composition of  claim 1 , wherein the in vitro pre-conditioned BMSC population are labeled with a bio-imaging agent. 
     
     
       8. A method of treating an enteric nervous system disorder comprising:
 administering to a subject in need thereof a pharmaceutical composition comprising an isolated in vitro pre-conditioned population of adult bone marrow derived mesenchymal stem cells (BMSCs), wherein at least 80% of the in vitro pre-conditioned BMSC population express PGP9.5 and NSE, 
 wherein at least 70% of the in vitro pre-conditioned BMSC population express Tuj1 and HuC/D, 
 wherein at least 75% of the in vitro pre-conditioned BMSC population express neuronal nitric oxide synthase (nNOS), and wherein the BMSCs promote de novo regeneration of neurons not originated from the BMSCs when the BMSCs are administered to a subject 
 and a pharmaceutically acceptable carrier. 
 
     
     
       9. The method of  claim 8 , wherein the enteric nervous system disorder is selected from the group consisting of a dysmotility syndrome, a diabetic gastroparesis, an intestinal pseudo-obstruction of motility, and neuronal loss in an enteric nervous system. 
     
     
       10. The method of  claim 8 , wherein the dysmotility syndrome is selected from the group consisting of achalasia, gastro-esophageal reflux disease, delayed emptying of the stomach, abdominal pain, bloating, diarrhea and constipation. 
     
     
       11. The method of  claim 10 , wherein the dysmotility syndrome is a congenital dysmotility syndrome. 
     
     
       12. The method of claim ll, wherein the congenital dysmotility syndrome is Hirschsprung disease. 
     
     
       13. The method of  claim 8 , wherein the pharmaceutical composition is administered into a gastrointenstinal tract of the subject. 
     
     
       14. The method of  claim 8 , wherein the pharmaceutical composition is administered endoscopically to the subject. 
     
     
       15. The method of  claim 8 , where the pharmaceutical composition is administered into a pylorus of the subject. 
     
     
       16. The method of  claim 15 , wherein the pharmaceutical composition is administered into a submucosal layer of the pylorus. 
     
     
       17. A method of preparing mesenchymal stem cells exhibiting a neuronal phenotype which method comprises:
 isolating mesenchymal stem cells from an adult bone marrow; 
 culturing the mesenchymal stem cells in a medium comprising glial cell derived neurotrophic factor and a fetal gut culture medium to thereby obtain an isolated in vitro pre-conditioned population of adult bone marrow derived mesenchymal stem cells (BMSCs), 
 wherein at least 80% of the in vitro pre-conditioned BMSC population express PGP9.5 and NSE, 
 wherein at least 70% of the in vitro pre-conditioned BMSC population express Tuj1 and HuC/D, 
 wherein at least 75% of the in vitro pre-conditioned BMSC population express neuronal nitric oxide synthase (nNOS), and 
 wherein the BMSCs promote de novo regeneration of neurons not originated from the BMSCs when the BMSCs are administered to a subject. 
 
     
     
       18. The method of  claim 17 , wherein the mesenchymal stem cells maintain the neuronal phenotype for at least 28 days in vivo. 
     
     
       19. The method of  claim 17 , wherein about 80% of the cells in the in vitro pre-conditioned BMSC population express PGP9.5, about 80% of the cells in the in vitro pre-conditioned BMSC population express NSE, about 75% of the cells in the in vitro pre-conditioned BMSC population express Tuj1, about 73% of the cells in the in vitro pre-conditioned BMSC population express HuC/D and about 75% of the in vitro pre-conditioned BMSC population express nNOS. 
     
     
       20. The method of  claim 17 , wherein the mesenchymal stem cells are human cells.